Accumulation of 2-deoxy-D-glucose-6-phosphate as a measure of glucose uptake in the isolated perfused heart: a 31P NMR study.
Accumulation of 2-deoxy-D-glucose-6-phosphate as a measure of glucose uptake in the isolated perfused heart: a 31P NMR study.
复制标题
2-脱氧-D-葡萄糖-6-磷酸的积累作为离体灌注心脏中葡萄糖摄取的测量：31P NMR 研究。
DOI：
10.1016/j.ymben.2003.10.002
复制
发表时间：
2004
影响因子：
8.4
通讯作者：
Clarke,Kieran
中科院分区：
文献类型：
--
作者：
Hopkins,JamesCA;Radda,GeorgeK;Veech,RichardL;Clarke,Kieran
2-脱氧-d-葡萄糖-6-磷酸（2DG 6P）的积累，使用31 P NMR光谱检测，已被用作葡萄糖摄取速率的测量，但该测量的准确性尚未得到验证。在这项研究中，离体大鼠心脏灌注不同的底物或异丙肾上腺素为30分钟，然后测量无论是2DG 6P积累或[2- 3 H]葡萄糖摄取，没有和有胰岛素。所有心脏的基础收缩功能和代谢物浓度均相同。2DG 6P蓄积的基础速率显著不同，取决于先前的灌注方案，并且是[2- 3 H]葡萄糖摄取速率的38-60%，而胰岛素刺激的2DG 6P蓄积与[2- 3 H]葡萄糖摄取速率相同或高71%。因此，2DG 6P蓄积/[2- 3 H]葡萄糖摄取速率的比值在0.38至1.71之间变化，这取决于先前的灌注条件或胰岛素的存在。2DG 6P的水解速率与细胞内2DG 6P浓度成正比，Km为17.5mM，Vmax为1.4μmol/g干重/min。我们认为2DG 6P的积累速率不能准确反映离体心脏在所有生理条件下的葡萄糖摄取速率，应谨慎使用。
The accumulation of 2-deoxy-d-glucose-6-phosphate (2DG6P), detected using31P NMR spectroscopy, has been used as a measure of the rate of glucose uptake, yet the accuracy of this measurement has not been verified. In this study, isolated rat hearts were perfused with different substrates or isoproterenol for 30min before measurement of either 2DG6P accumulation or [2-3H]glucose uptake, without and with insulin. Basal contractile function and metabolite concentrations were the same for all hearts. The basal rates of 2DG6P accumulation differed significantly, depending on the preceding perfusion protocol, and were 38–60% of the [2-3H]glucose uptake rates, whereas insulin-stimulated 2DG6P accumulation was the same or 71% higher than the [2-3H]glucose uptake rates. Therefore the ratio of 2DG6P accumulation/[2-3H]glucose uptake rates varied from 0.38 to 1.71, depending on the prior perfusion conditions or the presence of insulin. The rates of 2DG6P hydrolysis were found to be proportional to the intracellular 2DG6P concentrations, with a Kmof 17.5mM and Vmaxof 1.4μmol/g dry weight/min. We conclude that the rates of 2DG6P accumulation do not accurately reflect glucose uptake rates under all physiological conditions in the isolated heart and should be used with caution.